Fractionation of hydrogen isotopes in lipid biosynthesis

Isotopic compositions of carbon-bound hydrogen in individual compounds from eight di€erent organisms were measured using isotope-ratio-monitoring gas chromatography±mass spectrometry. This technique is capable of measuring D/H ratios at natural abundance in individual lipids yielding as little as 20 nmol of H2, and is applicable to a wide range of compounds including hydrocarbons, sterols, and fatty acids. The hydrogen isotopic compositions of lipids are controlled by three factors: isotopic compositions of biosynthetic precursors, fractionation and exchange accompanying biosynthesis, and hydrogenation during biosynthesis. dD values of lipids from the eight organisms examined here suggest that all three processes are important for controlling natural variations in isotopic abundance. n-Alkyl lipids are depleted in D relative to growth water by 113±262-, while polyisoprenoid lipids are depleted in D relative to growth water by 142±376-. Isotopic variations within compound classes (e.g., n-alkanes) are usually less than 050-, but variations as large as 150are observed among isoprenoid lipids from a single organism. Phytol is consistently depleted in D by up to 50relative to other isoprenoid lipids. Inferred isotopic fractionations between cellular water and lipids are greater than those indicated by previous studies. # 1999 Elsevier Science Ltd. All rights reserved.